Anti-Aspergillus spp. antibody (Ab) levels of seven mating pairs of captive African penguins (Spheniscus demersus) and the maternal Ab levels of their embryonated and unembryonated eggs were determined by ELISA. The egg Ab levels were significantly higher than the Ab levels of their female or male parents. Maternal Ab of unembryonated eggs and Ab of the embryos were significantly correlated with the Ab levels of their female parents. Because eggs can be sampled where adult penguins are not accessible, this technique can be applied for prediction of Aspergillus spp. Ab titers of penguin females in wild populations.